Method of increasing the diastase content of materials



1944- M. CHRISTENSEN METHOD OF INCREASING THE DIASTASE CONTENT OFMATERIALS Filed Oct. 50, 1941 2 Sheets-Sheet l SOYA MEAL MATERIAL HAVINGDIASTATIC PROPERTIES -SUCH AS BARLEY MALT, SOY BEANS OR GRINDING TOCOURSE IVIEALI IF NECESSARY PREFERABLY 0.2 N.

ADDING DIL'UTE HYDROCHLORIC ACID BY ,WEIGHT-pH 4.0 4.2

, MIXTURE WITH MOISTURE CONTENT OF 50% ASPERG/LLUS ORYZAE INCUBATING INTWO STAGES,

WITHOUT AND WITH AERATION BREAKING UP MATS FORMED IN INCUBATION FURTHERAERATION .PRODUCT HAVING INCREASED DIASTATIC PROPERTIES 3 m {to f!warrant/v 1 L. M. CHRISTENSEN 2,359,356

METHOD OF INCREASING THE DIASTASE CONTENT OF MATERIALS Filed Oct. 30,1941 2 Sheets-Sheet 2 F703 A MATERIAL HAVING DIASTATIC PROPERTIES SUCHAS BARLEY MALT, SOY BEANS OR SOYA MEAL MIXTURE WITH MOISTURE CONTENT OF50/ BYWEIGHT TREATING WITH A DIASTASE PRODUCING BACTERIA SUCH AS ONE OFTHE BACILLUS MESENTER/CUS GROUP f INCUBATION UNDER I AE oBID CONDITIONS7 I I I I I I ADDING LACTIC AcID I I PRODUCING BACTERIA ADDING DILUTE .7i HYDROCHLORIC ACI I D INCUBATING FOR APP- ROXIMATELY 24 HOURS I TOPRODUCE ACID MIXTURE OF pH 4.0-4.2

lNNOOULAT-ING WITH SPORES OF MOLD CAPABLE OF PRODUCING DIASTASE SUCH ASASPERGILLUS ORYZAE |INcuBATING IN 2 STAGES,WITHOUT AERATION FOLLOWED BYAERATIONJ I LBREAKING UP MATS 'FDRMEDEY INGUBATION awe/rm FURTHERAERATION [f0 fitf/B/Sff/VJf/V PRODUCT HAVING GREATLY IN- 9 1.2 4,,

cREAsED DI sTATIc PROPERTIES 6M zmea Patented Oct. 3, 1944 UNITED STATESPATENT OFFICE METHOD ormonmsmo 'rnn IIJIASTASE CONTENT OF MATERIALS LeoM. Christensen, Miller, Nelm, asslgnor to National Agrol Company, Inc.,New York, N. Y., a corporation of Delaware 1 Application October so,1941, Serial no. 417,206 8 Claims. (Cl. 195-73) This invention relatesto a method 01' enhancing the diastatic power of materials and moreespecially to a method .of increasing the diastatic power of materialsalready havinga diastase content.

The present application is a continuation in part of my co-pendingapplication entitled Methd of promoting mold growth, Serial No. 360,206,filed October 7, 1940.

One of the objects of the present invention is to provide a method ofenhancing thediastatic power of materials which already contain diastaseby inoculating the material with a diastase producing mold andsubjecting the mixture to conditions which will increase the diastati'cpower of the mixture.

A further'object of this invention is to provide a method of enhancingthe diastatic power ofmaterials which already contain diastase byinoculating the material with bacteria of a diastase producing typeandalso a diastase producing mold in order to provide a material havingan enhanced diastase content.

With these and other objects in view, the present invention embracesbroadly the idea of increasing the diastase content of material alreadyhaving diastatic power as for example soy beans, soya meal'or maltedcereal grain such as barley malt. In one method oi practicing theinvention, the selected material is inoculated with a mold capable ofproducingdiastase. In the preferred method, a diastase producingbacteria is added to the selected material and after allowing thebacteria to grow the diastatic mold is added and the material isincubated. In either process,'a product containing enhanced diastaticpower is produced.

Inthe drawings:

Figure l discloses a method of producing a material having enhanceddiastatic powers by adding a mold capable of producing diastase to aselected material having diastatic powers.

Figure 2 discloses a method in which a diastase producing bacteria isadded to the selected material and allowed to incubate before the moldspores are added. 4

In practicing the process" disclosed in Figure 1, a material havingdiastatic powers such as barley malt, soy beans, or soya meal from whichthe oil has been extracted is first selected and any solid matter whichmay be present is ground to form a coarse meal. Dilute hydrochloric acidis then added to this material in the proper proportions to form amixture having approximately 50% by weight 'of the moisture and ahydrogen ion content of 4.0 to 4.2. Ordinarily an acid solution of 2normal will be suitable to produce this result. The presence of, acidduring the process is necessary to prevent the growth of butyric acidproducing bacteria as the material cannot be sterilized withoutdestroying its diastase content. After the acid has been added, moldspores of a type capable of producing diastase are introduced in thesolution. Among these molds which are suitable for use in .the processmay be men-.

tioned Aspergillus oryzae and species of the genera Rhizopus and Mucor.The presence of the acid solution in the previously mentionedconcentration promotes the growth or these molds- The material, afterthe addition of the selected mold spores, is subjected'to an incubation.This incubation consists of approximately a 12 hour period withoutaeration and approximately a. 12 hour period in which the temperature ismaintained at less than 50 (7., and preferably between 40 and 45 C. bycontrolling the oxidatlve processes by aeration.

. After this treatment the mold mycelium ls iully grown and forms thematerial into a solid met. This mat is then broken into pieces ofroughlyone inch in diameter and the material is again aerated for approximately24 hours. The resulting product has a high diastase content as itconsists not only of the diastase contained in the origina1 material butalso that produced by the mold growth.

An apparatus which is suitable with slight changes to beused in thisprocess is disclosed in my eo-pendingapplication Serial No. 360,205,filed October 7, 1940, 'now Patent No. 2,325,368, issued July 27, 1943.

After the final aeration, the product may be dried and stored or itmaybe ground and used in a fermentation process to convert starchy materialinto carbohydrates. I1 desired water may be added and the product madeinto a slurry before it is added to the fermenation process.

In the process disclosed in Figure 2, sufllcient water is added to theselected material to form a moisture content which is 50% by weight ofthe total weight of the material. This mixture is then inoculated with abacteria of a type ca- .pable of producing diastase such as on'e\of theBacillus meseiztericus group.

After inoculation the bacteriaare allowed to grow for from 24 to 48hours under aerobic conditions produced by aeration to prevent thegrowth of butyric acid producing bacteria. II this aeration is notsufilcient to prevent this tic acid may be added in place of dilutehydrol0,

chloric acid in sufficient quantities to produce a pH of from 4.0 to 4.2before the mold spores are introduced into the mixture. This acid may beproduced by incubating a culture of lactic acid producing bacteria forapproximately 24 hours at a temperature of from 30 to 32 C. It is thenadded to the diastatic material.

After the proper hydrogen ion concentration has been produced by theaddition of the selected acid the process continues in the mannerpreviously described in connection with Figure 1. More specifically,after the addition of the selected mold spores the material is subjectedto the two stage incubation. As previously described, this incubationconsists of approximately a 12 hour period, without aeration andapproximately a 12 hour period in whichthe temperature is maintained atless than 50 C. preferablybetween 40 and 45 C. by controlling theoxidative processes by aeration.

After this treatment the mats formed by the growth of the mold myceliumare broken into pieces of roughly one inch in diameter and the materialis again aerated for approximately 24 hours. The resulting product has avery high diastase content due to the fact that it contains not only thediastase of the original starting material and that produced by the moldgrowth but also the diastase produced by the growth of the bacteria.

The final product may be dried and stored or it may be ground and usedin a fermentation process to convert starchy material to carbohydrates.If desired a slurry may be formed by adding water to the groundmaterial.

While for purposes of illustration, I have described two methods ofproducing a material of increased diastase content, it is obviousthat anumber of variations may be made without departing from the spirit ofthe invention. For example, other types of mold and bacteria of asuitable species may be employed, or the steps of th process may bevaried within certain well defined limits.

I claim:

1. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a diastatic material reducing thematerial to a coarse meal, adding a suihcient quantity of hydrochloricacid and water to the material to obtain a mixture having a pH ofapproximately 4.0 and a moisture content of approximately 50% by weight,adding spore of a diastatic mold to the mixture, incubating theinoculated material for a period without aeration, and then furtherincubating the material for an equal period of time with aeration toremove the heat or oxidation.

2. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a diastatic material reducing thematerial to a coarse meal, adding a suflicient quantity of hydrochloricacid and water to the material to obtain a mixture having a pH ofapproximately 4.0 and a moisture content of approximately 50% by weight,adding spores of a diastatic mold to the mixture, incubating theinoculated material for'a period of approximately twelve hours withoutaeration and then subjecting the material to a further incubation ofapproximately twelve hours with aeration to remove the heat ofoxidation.

3. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a diastatic material reducing thematerial to a coarse meal, adding a suflicient quantity of hydrochloricacid and water to the material to obtain a mixture having a pH withinthe range of from 4.0 to 4.2 and a moisture content of approximately 50%by weight, adding mold spores of the genus Aspergillus to the mixtureand then incubating the inoculated material for a period ofapproximately twelve hours without aeration and then subjecting thematerial to a further incubation of approximately twelve hours withaeration to remove the heat of oxidation.

4. A'method of preparing a diastatic material having an increaseddiastase content comprising selecting a diastatic material, grinding toform a coarse meal, adding a sufficient quantity of hydrochloric acidand water to the material to obtain a mixture having a pH within therange or from 4.0 to 4.2 and a moisture content of approximately 50% byweight, adding spores of a diastatic mold to the mixture, incubating theinoculated material for a period of approximately twelve hours withoutaeration, 'further incubating the material for a period of approximatelytwelve hours with aeration in order to maintain a temperature between 40and 50' C., breaking the mats formed during the incubation and thensubjecting the material to a further aeration.

5. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a maltedcereal grain, adding asumcient quantity of hydrochloric acid and water to the malted materialto obtain a mixture having a pH within the range of from 4.0 to 4.2 anda moisture content of approximately 50% by weight, adding mold spores ofthe genus Aspergillus to the mixture, incubating the inoculated materialfor a period of approximately twelve hours without aeration, furtherincubating the material for a period of approximately twelve hours withaeration to maintain a temperature of approximately 45 C., breaking themats formed during the incubation into small particles and thensubjecting the material to a further aeration.

6. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a diastatic material, addingsufficient water to obtain a mixture having a moisture content ofapproximately 50%, adding bacteria of the Bacillus mesentericus group,incubating .under aerobic conditions for from twenty-four to forty-eighthours, adding a sufficient quantity of hydrochloric acid to obtain a pHof approximately 4.0, inoculating with spores of a mold capable ofproducing diastase, incubating the inoculated material for a period ofapproximately twelve hours without aeration and then subjecting thematerial to a further incubation of appproximately twelve hours withaeration to remove the heat of oxidation.

7. A method of preparing a diastatic material having an increaseddiastase content comprising tent or approximate1y50%, adding bacteria ofthe Bacillus mesentericus group, incubating under aerobic conditions,adding a sufiiclent quantity of dilute hydrochloric acid to form a mixture having a pH within the range of from 4.0 to 4.2, inoculating withspores of the genus Aspergillus, incubating the inoculated material fora period of approximately twelve hours without aeration and thensubjecting the material to a further incubation of approximately twelvehours with aeration to remove the heat of oxidation.

8. A method of preparing a diastatic material having an increaseddiastase content comprising selecting a malted cereal grain grinding toform a coarse meal -adding sumcient water to obtain a mixture having amoisture content of approximately 50%, adding bacteria or the Bacillusmesentericus group, incubating under aerobic conditions for a period offrom twenty-four to fortyeight hours, adding a sufilcient quantity ofdilute hydrochloric acid to form a mixture having. a pH within the rangeof from 4.0 to 4.2, inoculating with spores of a mold capable ofproducing diastase, incubating the inoculated material for a period ofapproximately twelve hours without aeration and then subjecting thematerial to a further incubation of approximately twelve hours withaeration to remove the heat of oxidation.

LEO M. CI-IRIS'IENSEIT.

